Gene Editing in the Chagas Disease Vector Rhodnius prolixus by Cas9-Mediated ReMOT Control
Rhodnius prolixus is currently the model vector of choice for studying Chagas disease transmission, a debilitating disease caused by Trypanosoma cruzi parasites. However, transgenesis and gene editing protocols to advance the field are still lacking. Here, we tested protocols for the maternal delivery of CRISPR-Cas9 (clustered regularly spaced palindromic repeats/Cas-9 associated) elements to developing R. prolixus oocytes and strategies for the identification of insertions and deletions (indels) in target loci of resulting gene-edited generation zero (G0) nymphs. We demonstrate successful gene editing of the eye color markers Rp-scarlet and Rp-white, and the cuticle color marker Rp-yellow, with highest effectiveness obtained using Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) with the ovary-targeting BtKV ligand. These results provide proof of concepts for generating somatic mutations in R. prolixus and potentially for generating germ line-edited lines in triatomines, laying the foundation for gene editing protocols that could lead to the development of novel control strategies for vectors of Chagas disease.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:7 |
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| Enthalten in: |
The CRISPR journal - 7(2024), 2 vom: 19. Apr., Seite 88-99 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Lima, Leonardo [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 19.04.2024 Date Revised 19.04.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1089/crispr.2023.0076 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM370536959 |
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| 520 | |a Rhodnius prolixus is currently the model vector of choice for studying Chagas disease transmission, a debilitating disease caused by Trypanosoma cruzi parasites. However, transgenesis and gene editing protocols to advance the field are still lacking. Here, we tested protocols for the maternal delivery of CRISPR-Cas9 (clustered regularly spaced palindromic repeats/Cas-9 associated) elements to developing R. prolixus oocytes and strategies for the identification of insertions and deletions (indels) in target loci of resulting gene-edited generation zero (G0) nymphs. We demonstrate successful gene editing of the eye color markers Rp-scarlet and Rp-white, and the cuticle color marker Rp-yellow, with highest effectiveness obtained using Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) with the ovary-targeting BtKV ligand. These results provide proof of concepts for generating somatic mutations in R. prolixus and potentially for generating germ line-edited lines in triatomines, laying the foundation for gene editing protocols that could lead to the development of novel control strategies for vectors of Chagas disease | ||
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| 700 | 1 | |a Mota, Jamile |e verfasserin |4 aut | |
| 700 | 1 | |a Bressan, Daniel |e verfasserin |4 aut | |
| 700 | 1 | |a Julio, Alison |e verfasserin |4 aut | |
| 700 | 1 | |a Cavalcante, Robson |e verfasserin |4 aut | |
| 700 | 1 | |a Macias, Vanessa |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Zhiqian |e verfasserin |4 aut | |
| 700 | 1 | |a Rasgon, Jason L |e verfasserin |4 aut | |
| 700 | 1 | |a Bier, Ethan |e verfasserin |4 aut | |
| 700 | 1 | |a Araujo, Helena |e verfasserin |4 aut | |
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